Measurement of urinary estrogen metabolites using a monoclonal enzyme-linked immunoassay kit: assay performance and feasibility for epidemiological studies.

We evaluated an enzyme-linked immunoassay kit (Estramet 2/16) for the measurement of 2-hydroxyestrone (2-OH E1) and 16-alpha hydroxyestrone (16 alpha-OH E1), major metabolites of estradiol. Urine samples from 14 healthy premenopausal women on days 1, 8, 15, and 22 of their menstrual cycle were assayed along with standards, kit controls, and in-house controls. The intra-assay percentage CVs of 2-OH E1, 16 alpha-OH E1, and the 2-OH E1: 16 alpha-OH E1 ratio were 6.8, 7.4, and 1.8, respectively; the interassay percentage CVs were 15.3, 30.7, and 23.3, respectively. The assay linearity was between 0 and 40 ng/ml. The mean 2-OH E1:16 alpha-OH E1 ratio was relatively constant throughout the day, but it increased by around 50% between the follicular and luteal portions of the menstrual cycle. Individual reagent kits within each lot for 16 alpha-OH E1 were stable for 2 weeks. There was considerable lot-to-lot variation over a 5-month period. In lots used during the last 2 months of the study, values of 2-OH E1 from in-house controls increased by 30-50%, and those of 16 alpha-OH E1 by 50-100%, relative to values obtained initially on the same samples. Depending on the lot, the ratio of the two metabolites ranged from 2 to 5.5. These data suggest that the assay is useful for studies where samples can be assayed with the same kit lot over a period of not more than 2 weeks, but that it is not now suitable for studies that extend over a long enough period of time so that multiple kit lots are required.